Chemical Composition of Ballota macedonica Vandas and Ballota nigra L. ssp. foetida (Vis.) Hayek Essential Oils – The Chemotaxonomic Approach

The essential oils isolated from fresh aerial parts of Ballota macedonica (two populations) and Ballota nigra ssp. foetida were analyzed by GC and GC/MS. Eighty five components were identified in total; 60 components in B. macedonica oil (population from the Former Yugoslav Republic of Macedonia), 34 components in B. macedonica oil (population from the Republic of Serbia), and 33 components in the oil of B. nigra ssp. foetida accounting for 93.9%, 98.4%, and 95.8% of the total oils, respectively. The most abundant components in B. macedonica oils were carotol (13.7 – 52.1%), germacrene D (8.6 – 24.6%), and (E)‐caryophyllene (6.5 – 16.5%), while B. nigra ssp. foetida oil was dominated by (E)‐phytol (56.9%), germacrene D (10.0%), and (E)‐caryophyllene (4.7%). Multivariate statistical analyses (agglomerative hierarchical cluster analysis and principal component analysis) were used to compare and discuss relationships among Ballota species examined so far based on their volatile profiles. The chemical compositions of B. macedonica essential oils are reported for the first time.     

The toxicity of the Aggregatibacter actinomycetemcomitans cytolethal distending toxin correlates with its phosphatidylinositol‐3,4,5‐triphosphate phosphatase activity

The Aggregatibacter actinomycetemcomitans cytolethal distending toxin (Cdt) induces G2 arrest and apoptosis in lymphocytes and other cell types. We have shown that the active subunit, CdtB, exhibits phosphatidylinositol‐3,4,5‐triphosphate (PIP3) phosphatase activity, leading us to propose that Cdt toxicity is the result of PIP3 depletion and perturbation of phosphatidylinositol‐3‐kinase (PI‐3K)/PIP3/Akt signalling. To further explore this relationship, we have focused our analysis on identifying residues that comprise the catalytic pocket and are critical to substrate binding rather than catalysis. In this context, we have generated several CdtB mutants and demonstrate that, in each instance, the ability of the toxin to induce cell cycle arrest correlates with retention of phosphatase activity. We have also assessed the effect of Cdt on downstream components of the PI‐3K signalling pathway. In addition to depletion of intracellular concentrations of PIP3, toxin‐treated lymphocytes exhibit decreases in pAkt and pGSK3β. Further analysis indicates that toxin‐treated cells exhibit a concomitant loss in Akt activity and increase in GSK3β kinase activity consistent with observed changes in their phosphorylation status. We demonstrate that cell susceptibility to Cdt is dependent upon dephosphorylation and concomitant activation of GSK3β. Finally, we demonstrate that, in addition to lymphocytes, HeLa cells exposed to a CdtB mutant that retains phosphatase activity and not DNase activity undergo G2 arrest in the absence of H2AX phosphorylation. Our results provide further insight into the mode of action by which Cdt may function as an immunotoxin and induce cell cycle arrest in target cells such as lymphocytes.     

The Association Between Personality Traits,Morphological Traits and Alternative Mating Behaviour in Male Endler's Guppies,Poecilia wingei

Alternative mating behaviour, personality traits and morphological characters are predicted to be correlated. Bolder, larger and more colourful males are expected to preferentially court females, while shy, small and drab‐coloured individuals are predicted to sneak copulations. We used males of Endler's guppy, Poecilia wingei, to test this association over a long temporal period (hence including ontogenetic changes) and under two social environments (male‐biased and female‐biased). We found that personality traits (exploration, boldness, activity) of P. wingei males were highly repeatable across long time spans, but they were not correlated (formed no behavioural syndrome). Male age and social environment had no effect on any personality trait, despite their effects on alternative mating behaviour. Young males with higher activity levels were more likely to attempt sneaking. In older fish, there was an association between orange coloration, courtship and boldness, but this was not observed in young males. Our results suggest that alternative mating behaviour is more flexible than personality traits and is independent of them. Non‐colour‐based morphological traits (gonopodium length, body length, caudal straps length, dorsal fin length) were not correlated with any particular mating behaviour.     

Syntaxonomy of theTrifolio-Geranietea (saum vegetation) in Slovenia

This paper summarises present knowledge of the natural saum vegetation (Trifolio-Geranietea: Trifolion medii, Geranienion andDictamno-Ferulagenion) in Slovenia. The alliance ofGeranion sanguinei was subdivided into theGeranienio (primarily of Central European distribution) and SubmediterreanDictamno-Ferulagenion. Several new syntaxa (Scorzonero villosae-Trifolietum alpestris, Cirsio-Clematidetum rectae andOrigano-Cnidietum silaifolii) were described within theDictamno-Ferulagenion.     

Isolation and characterization of <Emphasis Type="Italic">Clostridium difficile</Emphasis> from shellfish and marine environments

This pilot study was carried out to evaluate the occurrence of Clostridium difficile in marine environments and in edible shellfish. Samples of seawater, sediment, and zooplankton were collected at five sampling stations in the Gulf of Naples. Six samples of edible shellfish, furthermore, were obtained: two from mussel farms and four from wholesalers. The isolation and the characterization of C. difficile strains were carried out using selective media and molecular techniques, respectively. C. difficile was isolated from nine of the 21 samples investigated. Shellfish and zooplankton showed the highest prevalence of positive samples. No C. difficile was detected in marine sediment. Majority of the C. difficile isolates were toxin A/B positive. Six known different PCR ribotypes (003, 005, 009, 010, 056, and 066) were identified, whereas one strain may represent a new PCR ribotype. C. difficile may be present in the marine environment in Southern Italy, including shellfish and zooplankton. This study is reporting the isolation of C. difficile from zooplankton, clams, and mussels and pointing out a new possible route to exposure to C. difficile of healthy individuals in the community.     

Epidemiologic characteristics of human campylobacteriosis in the County Primorsko-goranska (Croatia), 2003-2007

The aim of the study was to investigate campylobacteriosis incidence in the County Primorsko-goranska (Croatia) between 2003 and 2007 and to find out possible connection with environmental factors (the average monthly temperature and total monthly precipitation). The data (number of stool samples examined, age and sex distribution of patients, monthly distribution of isolates and distribution of isolates according to the species) from the Laboratory for Diagnostics of Enteric Infections of the Teaching Institute of Public Health of the County Primorsko-goranska (Croatia) were analyzed retrospectively. During the observed period 30,164 stool samples were examined for Campylobacter spp. Campylobacters were identified in 1,242 (4.12%) samples. The overall annual campylobacter incidence rate was 81.3 +/- 21.9/100,000 population. Campylobacter jejuni was found in 1,093 (88%) and C. coli in 149 (12%) patients. Our findings showed age distribution of patients typical for developed countries. The patients were mostly children under 5 years (484.4 +/- 129.1/ 100,000, p < 0.001) and between 5 and 9 years of age (226.5 +/- 60.5/100,000, p < 0.05). Male consistently experienced higher rates, but the difference between genders was significant in the age groups from birth till late twentieth (p < 0.001). Campylobacter rates were significantly associated with monthly average temperatures (p < 0.05), but not with precipitation. Further investigations into the incidence of campylobacteriosis on the national level are necessary. The causes of the noticed monthly distribution, sources of infection and connection with the routes of transmission in humans need to be elucidated as well.     

Advantages of intramedullar fixation in treatment of congenital tibial pseudoarthrosis--a case report

This report describes a case of a 29-year old patient with congenital pseudoarthrosis of the distal tibia previously treated unsuccessfully by a conventional surgical method. Tibial congenital pseudoarthrosis is a rare disease characterized by segmental osseous weakness resulting in deformation of the bone and spontaneous fractures which progresses to a tibial nonunion. In our case we used intramedullary stabilization with bone grafting and six month after operation congenital pseudarthrosis of the tibia healed.     

Age,growth and mortality of common two‐banded seabream,Diplodus vulgaris (Geoffroy Saint‐Hilaire, 1817), in the eastern Adriatic Sea (Croatian coast)

Age, growth and mortality were analysed for the common two‐banded seabream, Diplodus vulgaris, collected in the eastern Adriatic (Croatian coast) from commercial fishery catches by ‘tramata’ fishing (2005–2006) to obtain growth estimation. The oldest female was estimated to be age 11, the oldest male age 10 years. The von Bertalanffy growth parameters estimated by reading scales were: L_∞ = 48.60 cm (SE = 1.101), K = 0.112 (SE = 0.005) and t₀ = ?2.366 (SE = 0.060) for all specimens; L_∞ = 51.96 cm (SE = 2.153), K = 0.095 (SE = 0.007) and t₀ = ?2.837 (SE = 0.120) for females and L_∞ = 56.25 cm (SE = 2.662), K = 0.084 (SE = 0.067) and t₀ = ?2.920 (SE = 0.117) for males. The overall sex ratio was 1.22 : 1 in favour of males. Total mortality, corresponding to the slope of the descending limb of the catch curve, was Z = 0.81 per year for females and Z = 0.85 per year for males. Exploitation ratios were E = 0.68 for females and E = 0.73 for males.     

Incidence and factors associated with early pregnancy losses in Simmental dairy cows

It has been suggested that management system, milk yield, parity, body condition score and ambient temperature can significantly influence the rate of early pregnancy loss in dairy cattle. The objectives of this study were to establish the extent and patterns of early pregnancy loss from days 32 to 86 of gestation, and to check relationships between management system, milk yield, ambient temperature (quartile), body condition score, bull and parity on the early pregnancy loss rate for Simmental dairy cattle in Croatia. Animals were housed in two dairy farms with two different management systems (pasture based-group A, n=435 and intensive-group B, n=425) with a total of 151 heifers and 709 cows. Overall pregnancy losses were recorded in 67 (7.79%) animals, with late embryonic losses in 30 (44.77%) and early fetal losses in 37 (55.23%) animals (P>0.05). Early pregnancy losses were twofold higher in group B when compared to the group A (P<0.05). More than the half of pregnancy losses were recorded during the III quartile (P<0.05). There was no significant relationship between the paternal bull and pregnancy loss rate. Low body condition score (BCS 2-3) was associated with the highest, while BCS 3.25-4 showed the lowest pregnancy loss rate (P<0.05). The pregnancy loss rate increased in parallel with parity and milk yield increase.